Practical Preimplantation Genetic Diagnosis

Practical Preimplantation Genetic Diagnosis

Yury Verlinsky and Anver Kuliev

Practical

Preimplantation Genetic Diagnosis

With 125 Figures

Yury Verlinsky, PhD Anver Kuliev, MD, PhD

Reproductive Genetics Institute, Chicago, IL, USA

British Library Cataloguing in Publication Data

A catalogue record for this book is available from the British Library

Library of Congress Control Number: 2005923550

ISBN-10: 1-85233-920-9 Printed on acid-free paper ISBN-13: 978-1-85233-920-3

C Springer-Verlag London Limited 2005

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Contents

Foreword . . . . vii

Preface . . . . ix

Acknowledgements . . . . xi

Contributors . . . . xiii

1. Primary Prevention of Genetic Disorders and Place of Preimplantation Genetic Diagnosis . . . . 1

2. Approaches to Preimplantation Genetic Diagnosis . . . . 7

2.1 Obtaining Biopsy Material . . . . 8

2.1.1 Polar Body Sampling . . . . 9

2.1.2 Preconception Testing for Paternally Derived Mutations by Sperm Duplication . . . . 14

2.1.3 Development of Artificial Human Gametes in Vitro . . . . 14

2.1.4 Embryo Biopsy . . . . 16

2.2 Single-Cell Genetic Analysis . . . . 17

2.2.1 DNA Analysis . . . . 17

2.2.2 FISH Analysis . . . . 20

3. preimplantation Genetic Diagnosis for Single Gene Disorders . . . . 29

3.1 Autosomal Recessive Diseases . . . . 29

3.2 Autosomal Dominant Disorders . . . . 41

3.2.1 Primary Torsion Dystonia . . . . 41

3.2.2 Charcot-Marie-Tooth Disease . . . . 48

3.4 Homozygous or Double Heterozygous Recessive Conditions . . . . 55

3.5 Conditions with No Available Direct Mutation Testing . . . . 59

3.6 Late-Onset Disorders with Genetic Predisposition . . . . 63

3.6.1 p53 Tumour-Suppressor Gene Mutations . . . . 66

3.6.2 Neurofibromatosis . . . . 69

3.6.3 Other Cancers . . . . 74

3.6.4 Alzheimer Disease . . . . 83

3.7 Blood Group Incompatibility . . . . 86

vi CONTENTS

3.8 Congenital Malformations . . . . 89

3.8.1 Sonic Hedgehog Mutation . . . . 90

3.8.2 Currarino Syndrome . . . . 93

3.8.3 Crouzon Syndrome . . . . 99

3.9 Dynamic Mutations . . . . 100

4. Preimplantation HLA Typing . . . . 111

4.1 PGD for Fanconi Anemia with Preimplantation HLA Matching . . . . 111

4.2 Current Experience of PGD with Preimplantation HLA Matching . . . . 113

4.3 Preimplantation HLA Matching Without PGD . . . . 124

4.4 Worldwide Experience on Preimplantation HLA Typing . . . . 128

5. Preimplantation Genetic Diagnosis for Chromosomal Disorders . . . . 135

5.1 Aneuploidies . . . . 140

5.1.1 Complex Errors and Aneuloidy Rescue in Female Meiosis . . . . 146

5.1.2 Mitotic Errors in Cleaving Embryos in Relation to Meiosis Errors . . . . 149

5.1.3 PCR-Based Aneuploidy Testing in Cleaving Embryos . . . . 152

5.1.4 Practical Relevance of Autosomal Monosomy Detection . . . . 153

5.1.5 Uniparental Disomies . . . . 155

5.2 Chromosomal Rearrangements . . . . 157

6. Clinical Outcomes Following Preimplantation Genetic Diagnosis . . . . 169

7. Preimplantation Genetic Diagnosis and Establishment of Human Embryonic Stem Cell Lines . . . . 175

7.1 Marfan Syndrome . . . . 179

7.2 Thalassemia . . . . 179

7.3 Fragile-X Syndrome . . . . 179

7.4 Myotonic Dystrophy . . . . 182

7.5 Neufibromatosis Type 1 . . . . 182

7.6 Becker Muscular Dystrophy . . . . 182

8. Ethical, Social, and Legal Issues . . . . 189

Index . . . . 195

Foreword

The casual reader may not fully appreciate the significance of the well-chosen title for this unique and welcome volume, Practical Preimplantation Genetic Diagnosis. The title is not only apropos but provided by the team that truly made it possible.

Prenatal genetic diagnosis began in the 1950s, with detection of X-chromatin in amniotic fluid cells. By the late 1960s amniocentesis allowed diagnosis of chromosomal abnormalities and selected inborn errors of metabolism. Labs sprung up worldwide, and in the 1970s antenatal testing became standard in certain patients. The 1980s brought two key advances.

First was noninvasive screening, with an invasive procedure like amniocentesis performed only if the risk was high enough to be justified. Concurrently, there was movement toward prenatal diagnosis earlier in pregnancy, as witnessed by chorionic villus sampling (CVS).

Still, diagnosis was not possible before 10 to 12 weeks, and, hence, clinical pregnancy termination was still necessary.

Although proof of the principle for preimplantation genetic diagnosis (PGD) had been shown in 1967 by Professor Robert Edwards through X-chromatin analysis in rabbit blas- tocysts, application in the human awaited advances in human assisted reproductive tech- nologies (ARTs) and in molecular techniques. The first human PGD finally came only in 1990. During the next few years, there was great interest in PGD in the genetic commu- nity; however, relatively few cases were performed, mostly for mendelian disorders. With introduction of fluorescent in situ hybridization (FISH), however, PGD was burgeoning.

By 2000, novel indications were added, including aneuploidy testing to identify chromo- somally normal embryos. Indeed, this is now the most common indication for PGD.

Practical Preimplantation Genetic Diagnosis systematically covers indications and tech-

nology. One is taken through PGD from start (embryo) biopsy to end (consequences), and

we learn from the group that has performed half the world’s cases. The technique for obtain-

ing embryonic DNA is first described (polar body biopsy or blastomere biopsy), followed

by single-cell genetic analysis (FISH or polymerase chain reaction–based approaches). The

spectrum of mendelian disorders and their detection follows. Once controversial topics like

late-onset disorders are covered, with especially lucid coverage on PGD to obtain human

leukocyte antigen (HLA)-compatible embryos. Preimplantation genetic diagnosis not only

avoids another affected offspring, but also provides umbilical cord–derived stem cells for

transplantation into an older moribund sib. Both numerical and structural chromosomal

abnormalities are discussed. We are given novel information on the unexpectedly high

sequential error rate in meiosis I followed by meiosis II. Clinical outcome and the safety

of PGD are discussed (and reassurance is given). Throughout, the discussion flows seam-

lessly between indication and counseling (reflecting the impeccable genetic credentials of

the authors) and laboratory performance (likewise). Illustrations are highly informative

and lucid. This is invaluable in understanding PGD for mendelian disorders, the diagnosis

of which requires linkage analysis. The illustrations make a potentially arcane topic quite

clear.

viii FOREWORD

Overall, one comes away with a keen appreciation of how PGD has moved from “bou- tique” medicine, practiced only rarely by the cognoscenti, to a technique rapidly becoming de rigueur in all in vitro fertilization (IVF) centers. Within 5 to 10 years, surely most, if not all, ART will require genetic testing prior to embryo transfer.

Considerable information is new and not heretofore available even in peer review format.

The result is a pleasing combination of reading a textbook peppered with peer review information. The sense of movement in PGD is thus palpable.

Practical Preimplantation Genetic Diagnosis is thus a gem. In this age of edited works, a text written by only one or two authors is rare. That our authors practically invented the field, and certainly have made it practical, makes this volume truly extraordinary.

This volume should thus be at the reach of every reproductive geneticist, ART lab, and reproductive biologist. The avid student will come away fully informed and up to date, and also ready for tomorrow’s advances.

Joe Leigh Simpson, M.D.

Ernst W. Bertner Chairman and Professor

Department of Obstetrics and Gynecology

Houston, Texas

June 2005

Preface

Although treatment is the major goal in the control of genetic disease, as in other fields of medicine, this is not yet a reality for most inherited conditions. Even with dramatic advancement in the field of gene therapy, there are not, unfortunately, many success stories to allow predicting its sound impact in the near future. Therefore, in the absence of radical treatment, prevention of genetic disorders is still one of the available options for genetically disadvantaged people. Among the measures for prevention, the avoidance of the birth of an affected child to couples at genetic risk has become a quite acceptable option in many populations. This is based on population screening and prenatal diagnosis, and has been quite successful, resulting in almost eradicating new cases of some genetic diseases from several Mediterranean populations, such as those in Sardinia and Cyprus. However, this has generated an increasing number of abortions following prenatal diagnosis, leading to a growing concern and negative reaction in society to the preventive genetics programs.

For example, some ethnic groups cannot accept any control measures regarding congenital diseases because pregnancy termination is not allowed due to social or religion reasons.

Even in those communities where abortion is allowed, some couples have to experience two or more terminations of pregnancies before they can have a normal child. No doubt these families might require an alternative option to achieve their desired family size without the need for termination of pregnancy.

The present book is devoted to a principally new approach in prevention of genetic disorders, which avoids the need for prenatal diagnosis and termination of pregnancy.

Accordingly, a novel concept of prepregnancy diagnosis is introduced, which is based on the control of the processes of the oocyte maturation, fertilization, and implantation, so to select and transfer back to patients only normal embryos, and achieve an unaffected preg- nancy resulting in the birth of a healthy child. In this way, the couples at high risk of having an offspring with genetic disease have an option to control the outcome of their pregnancy from the onset. So the place of this approach in the context of other available approaches for prevention of genetic disorders is discussed, together with other primary preventive measures, which may allow presently avoiding up to a half of congenital malformations presented at birth (Chapter 1). Although the option of prepregnancy diagnosis involves ovarian stimulation and in vitro fertilization (IVF), the description of the available expe- rience demonstrates that this has appeared to be an acceptable procedure in many ethnic groups all over the world.

As described in the methodological section (Chapter 2), preimplantation genetic di-

agnosis (PGD) may be achieved by testing of the female, and perhaps in the future also

male, gametes or single cells from the preimplantation embryo. Both of these methods have

been used in clinical practice and have shown to be useful in prediction of the genotype

of the resulting pregnancy, although many problems still remain to be resolved, requir-

ing careful consideration. The experience in the application of both of these approaches

and the progress in overcoming the existing problems are described, based on the original

practice of more than 3500 PGD cycles, presently comparable to the overall available world

x PREFACE

experience, as well as the experiences from the other active PGD centres. Because basic clinical methods for performing PGD are similar to those in IVF, which have been de- scribed previously in detail, the emphasis in this book is mainly on the laboratory aspects.

However, the clinical impact of PGD, and some of the ethical, social, and legal aspects are also briefly explored (Chapters 6 and 8). The present progress of PGD makes clear that the at-risk couples have to be aware of all available preventive options, including PGD, which is a complement rather than an alternative to the available preventive genetics services, allowing the couples to make their own choices.

Because PGD of single-gene disorders involves extensive preparatory work for each particular condition, a special section is devoted to the description of the PGD strategy for each group of these disorders. This includes the description of primer sequences, PCR conditions, polymorphic markers, the experience of the application of conventional and fluorescence PCR, allele drop out (ADO) rates, and other expected problems leading to misdiagnosis, with the detailed presentation of relevant case reports (Chapter 3). A special section is also devoted to preimplantation nongenetic testing, involving HLA typing, which has recently become one of the most attractive PGD applications allowing improved access to HLA-compatible stem cell transplantation (Chapter 4).

The application of PGD is of particular interest for assisted reproduction practices, so this is described in detail in a special section, providing strong evidence of PGD impact on pregnancy outcome (Chapters 5 and 6). This is not surprising, as the introduction of methods for preselection of cytogenetically normal and avoiding the transfer of cytoge- netically abnormal embryos should considerably improve the efficiency of IVF, based on the fact that aneuploidies are major contributors to spontaneous abortions and may also explain some of the implantation failures. Although more data are still needed to con- firm positive impact on birth outcome, an extensive original experience of FISH analysis of over 15,000 oocytes and embryos is presented (Chapter 5), together with the analysis of previous reproductive histories of patients, demonstrating strong evidence of clinical impact (Chapter 6). These chapters also include the follow-up studies of detected cases with chromosomal abnormalities in the different stages of preimplantation development to explore the problems of mosaicism and its biological significance at the cleavage stage.

It is presently being further confirmed that PGD is the most attractive option for cou- ples carrying balanced translocations (Chapter 5). In light of these data, a pioneering work on conversion of interphase nuclei of single biopsied blastomeres into metaphase chro- mosomes, based on the use of the modern nuclear transfer techniques, is described with presentation of the original experience of the application of these methods for PGD of translocations.

For the first time, the original experience of the latest applications of PGD to embryonic stem cells is described, providing the possibility of obtaining preimplantation embryos with known genotypes as a source for the establishment of custom-made embryonic stem cells (Chapter 7).

Finally, the current status of PGD is summarized, with prospects of the application of

PGD to medical practice. This information may help not only in planning and organiza-

tion of such services, but also in providing a working manual for the establishment and

performance of PGD in the framework of IVF and genetic practices.

Acknowledgements

We are indebted to our colleagues in DNA, FISH, and embryology laboratories, O. Verlinsky,

T. Sharapova, S. Ozen, K. Lazyuk, G. Wolf, Y. Illkevitch, N. Strelchenko, V. Galat,

V. Kuznyetsov, Z. Zlatoposlky, M. Chmura, V. Koukharenko, and I. Kirillova, and also

to our genetic counsellors, C. Lavine, R. Beck, R. Genoveze, and D. Pauling for their par-

ticipation in acquisition of the data and technical assistance.

Contributors

Svetlana Rechitsky, PhD, Single-Gene Disorders and HLA Typing

Jeanine Cieslak, MLT,

Micromanipulation and Chromosomal

Disorders